Inhibition of Encephalomyocarditis Virus and Poliovirus Replication by Quinacrine: Implications for the Design and Discovery of Novel Antiviral Drugs

Gasparian, Alexander V.; Neznanov, Nickolay; Jha, Sujata; Galkin, A. Yu.; Moran, John J.; Gudkov, Andrei V.; Gurova, Katerina V.; Komar, Anton A.

doi:10.1128/jvi.02569-09

Cited by 35 publications

(32 citation statements)

References 44 publications

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“…Nucleic acid-intercalating drugs were shown to inhibit IRES-mediated translation more than cap-dependent translation at low concentrations (10 to 20 M) (31). It was subsequently shown that QC inhibited the IRES activities of encephalomyocarditis virus (EMCV), hepatitis C virus (HCV), and poliovirus in a dose-dependent manner by interacting with the highly complex secondary structures of their IRES regions (32). The same study also found that the cellular p53 IRES, which has a much less complex secondary structure, is not sensitive to QC (32).…”

Section: Wssv Icp35 Is a Highly Expressed Nonstructural Proteinsupporting

confidence: 51%

“…10B). QC is presumably able to do this by virtue of its high affinity of binding to relatively complex secondary and/or tertiary structures (32). Unlike most other DNA viruses, WSSV has many gene clusters, and since these are all likely to use IRES elements to regulate translation, the same mechanism would also inhibit the translation of other important WSSV proteins encoded by these clusters.…”

Section: Discussionmentioning

confidence: 99%

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The DNA Virus White Spot Syndrome Virus Uses an Internal Ribosome Entry Site for Translation of the Highly Expressed Nonstructural Protein ICP35

Kang

Wang

Yang

et al. 2013

J Virol

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Although shrimp white spot syndrome virus (WSSV) is a large double-stranded DNA virus (ϳ300 kbp), it expresses many polycistronic mRNAs that are likely to use internal ribosome entry site (IRES) elements for translation. A polycistronic mRNA encodes the gene of the highly expressed nonstructural protein ICP35, and here we use a dual-luciferase assay to demonstrate that this protein is translated cap independently by an IRES element located in the 5= untranslated region of icp35. A deletion analysis of this region showed that IRES activity was due to stem-loops VII and VIII. A promoterless assay, a reverse transcription-PCR together with quantitative real-time PCR analysis, and a stable stem-loop insertion upstream of the Renilla luciferase open reading frame were used, respectively, to rule out the possibility that cryptic promoter activity, abnormal splicing, or read-through was contributing to the IRES activity. In addition, a Northern blot analysis was used to confirm that only a single bicistronic mRNA was expressed. The importance of ICP35 to viral replication was demonstrated in a double-stranded RNA (dsRNA) interference knockdown experiment in which the mortality of the icp35 dsRNA group was significantly reduced. Tunicamycin was used to show that the ␣ subunit of eukaryotic initiation factor 2 is required for icp35 IRES activity. We also found that the intercalating drug quinacrine significantly inhibited icp35 IRES activity in vitro and reduced the mortality rate and viral copy number in WSSV-challenged shrimp. Lastly, in Sf9 insect cells, we found that knockdown of the gene for the Spodoptera frugiperda 40S ribosomal protein RPS10 decreased icp35 IRES-regulated firefly luciferase activity but had no effect on cap-dependent translation.

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Section: Wssv Icp35 Is a Highly Expressed Nonstructural Proteinsupporting

confidence: 51%

Section: Discussionmentioning

confidence: 99%

The DNA Virus White Spot Syndrome Virus Uses an Internal Ribosome Entry Site for Translation of the Highly Expressed Nonstructural Protein ICP35

Kang

Wang

Yang

et al. 2013

J Virol

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“…S2). Subsequently, several compounds were used to validate these results: the anthracycline analogues DNR and EPI, the non-anthracyclines etoposide and ellipticine, and the known EV IRES inhibitor quinacrine (Gasparian et al, 2010;Wang et al, 2013) were used to treat the cells transfected with the dicistronic reporter mRNAs. The cells treated with the anthracycline analogues and quinacrine exhibited similar trends: decreasing levels of the EV IRES activities and statistically unaltered p53 IRES activities (Fig.…”

Section: Mechanism-of-action Of Idrmentioning

confidence: 99%

“…A fixed concentration of IDR (4 mM) was titrated with the EV 59 UTR RNAs and cellular p53 IRES RNA samples at 0.25-1 mM, and the fluorescence intensities were measured. Poly(rU) was used as a negative control because it is known to be unstructured with limited binding to the known nucleic acid ligands (Chaires, 2005;Gasparian et al, 2010), whereas a highly polymerized, long-stretch calf thymus DNA that efficiently binds to IDR was used as a positive control (Chaires, 2005;Charak & Mehrotra, 2013;Ozluer & Kara, 2014). EMCV 59 UTR RNA was also included for comparison.…”

Section: Idr Displays Stronger Affinities To the Ev 59 Utr Rnasmentioning

confidence: 99%

Idarubicin is a broad-spectrum enterovirus replication inhibitor that selectively targets the virus internal ribosomal entry site

Hou¹,

Lü

Wu³

et al. 2016

Journal of General Virology

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Enterovirus 71 (EV71) causes life-threatening diseases with neurological manifestations in young children. However, the treatment of EV71 infections remains an unmet medical need. Idarubicin (IDR) is an anthracycline compound that is used therapeutically for certain types of tumour. In this study, we identified IDR as an EV71 inhibitor, which displayed antiviral potency in the submicromolar range and substantially protected cells from the cytopathic effects and cell death caused by EV71 infections. The antiviral effects extended to several other enterovirus (EV) species, and these effects were independent of cytotoxicity or topoisomerase inhibition. Structure-activity relationship studies indicated the importance of the anthracycline scaffold for anti-EV potency. IDR effectively blocked the synthesis of viral protein and RNA, but not the viral proteolysis processes. Moreover, anthracyclines were demonstrated to suppress EV internal ribosomal entry site (IRES)-mediated translation; conversely, the cellular p53 IRES activity was not sensitive to IDR action. Inhibition of IRES-mediated translation by IDR correlated with the affinity of binding between IDR and the particular IRES. Moreover, IDR impaired binding between the EV71 IRES RNA and hnRNP A1, a known host IRES trans-acting factor. In sum, we have identified a USA FDA-approved anticancer drug with the new indication as a selective EV IRES binder and inhibitor. The finding may also provide leads for the development of novel antiviral therapies directed at the EV IRES RNA.

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“…21,22 On the other hand, the nucleic acid-binding ligand acriflavine inhibits protein synthesis in cell free lysates with specificity for internal initiation of translation. 23 However, the efficacy of most these molecules in animal models are not well documented and their mode of action are still poorly understood.…”

Section: Introductionmentioning

confidence: 99%

Local RNA flexibility perturbation of the IRES element induced by a novel ligand inhibits viral RNA translation

et al. 2015

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(2015) Local RNA flexibility perturbation of the IRES element induced by a novel ligand inhibits viral RNA translation, RNA Biology, 12:5, 555-568, DOI: 10.1080/15476286.2015 The internal ribosome entry site (IRES) element located at the 5´untranslated genomic region of various RNA viruses mediates cap-independent initiation of translation. Picornavirus IRES activity is highly dependent on both its structural organization and its interaction with host factors. Small molecules able to interfere with RNA function are valuable candidates for antiviral agents. Here we show that a small molecule based on benzimidazole (IRAB) inhibited foot-andmouth disease virus (FMDV) IRES-dependent protein synthesis in cells transfected with infectious RNA leading to a decrease of the virus titer, which was higher than that induced by a structurally related benzimidazole derivative. Interestingly, IRAB preferentially inhibited IRES-dependent translation in cell free systems in a dose-dependent manner. RNA structural analysis by SHAPE demonstrated an increased local flexibility of the IRES structure upon incubation with IRAB, which affected 3 stem-loops (SL) of domain 3. Fluorescence binding assays conducted with individual aminopurinelabeled oligoribonucleotides indicated that the SL3A binds IRAB (EC 50 18 mM). Taken together, the results derived from SHAPE reactivity and fluorescence binding assays suggested that the target site of IRAB within the FMDV IRES might be a folded RNA structure that involves the entire apical region of domain 3. Our data suggest that the conformational changes induced by this compound on a specific region of the IRES structure which is essential for its activity is, at least in part, responsible for the reduced IRES efficiency observed in cell free lysates and, particularly, in RNA-transfected cells.

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Inhibition of Encephalomyocarditis Virus and Poliovirus Replication by Quinacrine: Implications for the Design and Discovery of Novel Antiviral Drugs

Cited by 35 publications

References 44 publications

The DNA Virus White Spot Syndrome Virus Uses an Internal Ribosome Entry Site for Translation of the Highly Expressed Nonstructural Protein ICP35

The DNA Virus White Spot Syndrome Virus Uses an Internal Ribosome Entry Site for Translation of the Highly Expressed Nonstructural Protein ICP35

Idarubicin is a broad-spectrum enterovirus replication inhibitor that selectively targets the virus internal ribosomal entry site

Local RNA flexibility perturbation of the IRES element induced by a novel ligand inhibits viral RNA translation

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